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首页> 外文期刊>Journal of chromatography, A: Including electrophoresis and other separation methods >Characterization and identification of pradimicin analogs from Actinomadura hibisca using liquid chromatography-tandem mass spectrometry
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Characterization and identification of pradimicin analogs from Actinomadura hibisca using liquid chromatography-tandem mass spectrometry

机译：液相色谱-串联质谱法表征和鉴定猕猴桃中的菊苣素类似物

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摘要

Microbial cultures produce complex and potentially interesting mixtures of biosynthetic intermediates and derivatives of metabolites. These mixtures' reliable identification is important and so too is the development of techniques for their analysis. Here, a simple and highly selective method of detecting the biosynthetic congeners involved in the pentangular polyphenol pradimicin (PR) pathway from Actinomadura hibisca fermentation was developed. Solid-phase extraction (SPE) cleanup using an OASIS HLB cartridge was a simple and reliable tool for the extraction of PRs from a fermentation broth. The separation of each natural PR analog - eluted with a gradient system of aqueous acetonitrile through a reversed-phase C_(18) column containing ammonium acetate and acetic acid as additives - allowed their simultaneous profiling. The combined use of SPE cleanup and chromatographic separation, coupled with electrospray ionization-tandem mass spectrometry (ESI-MS/MS) detection was demonstrated to be sufficiently accurate and reliable to analyze the natural PR analogs produced from A. hibisca. Ten natural PRs were identified: four alanine-containing (PRA, PRC, PRL, and PRB), two glycine-substituted (PRD and PRE), and four serine-substituted (PRFA~(-1), PRFA-2, PRFL, and PRFB). This report demonstrates the first use of both SPE cleanup and HPLC-ESI-MS/MS to profile a wide range of structurally closely related PRs in a bacterial fermentation broth.
机译：微生物培养产生生物合成中间体和代谢物衍生物的复杂且可能令人感兴趣的混合物。这些混合物的可靠识别非常重要，分析技术的发展也很重要。在这里，开发了一种简单且高度选择性的方法，用于检测来自Actinomadura hibisca发酵的矩形多酚Pradimicin（PR）途径的生物合成同源物。使用OASIS HLB滤芯进行固相萃取（SPE）净化是从发酵液中提取PR的简单可靠的工具。分离每种天然PR类似物-通过乙腈水溶液的梯度系统通过含有乙酸铵和乙酸作为添加剂的反相C_（18）色谱柱进行洗脱-可以同时进行分析。 SPE净化和色谱分离的结合使用，再结合电喷雾电离串联质谱（ESI-MS / MS）检测，被证明是足够准确和可靠的，可以分析由木槿产的天然PR类似物。确定了十个天然PR：四个含丙氨酸的（PRA，PRC，PRL和PRB），两个甘氨酸取代的（PRD和PRE）和四个丝氨酸取代的（PRFA〜（-1），PRFA-2，PRFL，和PRFB）。该报告证明了SPE净化和HPLC-ESI-MS / MS的首次使用，可对细菌发酵液中各种结构密切相关的PR进行分析。

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